Upregulated ribosome pathway plays a key role in HDAC4, improving the survival rate and biofunction of chondrocytes
Aims: To explore the novel molecular mechanisms of histone deacetylase 4 (HDAC4) in chondrocytes via RNA sequencing (RNA-seq) analysis. Methods: Empty adenovirus (EP) and a HDAC4 overexpression adenovirus were transfected into cultured human chondrocytes. The cell survival rate was examined by real-...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
The British Editorial Society of Bone & Joint Surgery
2023-07-01
|
| Series: | Bone & Joint Research |
| Subjects: | |
| Online Access: | https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.127.BJR-2022-0279.R2 |
| _version_ | 1827897250474360832 |
|---|---|
| author | Li Guo Hua Guo Yuanyu Zhang Zhi Chen Jian Sun Gaige Wu Yunfei Wang Yang Zhang Xiaochun Wei Pengcui Li |
| author_facet | Li Guo Hua Guo Yuanyu Zhang Zhi Chen Jian Sun Gaige Wu Yunfei Wang Yang Zhang Xiaochun Wei Pengcui Li |
| author_sort | Li Guo |
| collection | DOAJ |
| description | Aims: To explore the novel molecular mechanisms of histone deacetylase 4 (HDAC4) in chondrocytes via RNA sequencing (RNA-seq) analysis. Methods: Empty adenovirus (EP) and a HDAC4 overexpression adenovirus were transfected into cultured human chondrocytes. The cell survival rate was examined by real-time cell analysis (RTCA) and EdU and flow cytometry assays. Cell biofunction was detected by Western blotting. The expression profiles of messenger RNAs (mRNAs) in the EP and HDAC4 transfection groups were assessed using whole-transcriptome sequencing (RNA-seq). Volcano plot, Gene Ontology, and pathway analyses were performed to identify differentially expressed genes (DEGs). For verification of the results, the A289E/S246/467/632 A sites of HDAC4 were mutated to enhance the function of HDAC4 by increasing HDAC4 expression in the nucleus. RNA-seq was performed to identify the molecular mechanism of HDAC4 in chondrocytes. Finally, the top ten DEGs associated with ribosomes were verified by quantitative polymerase chain reaction (QPCR) in chondrocytes, and the top gene was verified both in vitro and in vivo. Results: HDAC4 markedly improved the survival rate and biofunction of chondrocytes. RNA-seq analysis of the EP and HDAC4 groups showed that HDAC4 induced 2,668 significant gene expression changes in chondrocytes (1,483 genes upregulated and 1,185 genes downregulated, p < 0.05), and ribosomes exhibited especially large increases. The results were confirmed by RNA-seq of the EP versus mutated HDAC4 groups and the validations in vitro and in vivo. Conclusion: The enhanced ribosome pathway plays a key role in the mechanism by which HDAC4 improves the survival rate and biofunction of chondrocytes. Cite this article: Bone Joint Res 2023;12(7):433–446. |
| first_indexed | 2024-03-12T22:46:30Z |
| format | Article |
| id | doaj.art-c4ac13bff6184c008a47bea7f98f73b5 |
| institution | Directory Open Access Journal |
| issn | 2046-3758 |
| language | English |
| last_indexed | 2024-03-12T22:46:30Z |
| publishDate | 2023-07-01 |
| publisher | The British Editorial Society of Bone & Joint Surgery |
| record_format | Article |
| series | Bone & Joint Research |
| spelling | doaj.art-c4ac13bff6184c008a47bea7f98f73b52023-07-21T05:13:20ZengThe British Editorial Society of Bone & Joint SurgeryBone & Joint Research2046-37582023-07-0112743344610.1302/2046-3758.127.BJR-2022-0279.R2Upregulated ribosome pathway plays a key role in HDAC4, improving the survival rate and biofunction of chondrocytesLi Guo0Hua Guo1Yuanyu Zhang2Zhi Chen3Jian Sun4Gaige Wu5Yunfei Wang6Yang Zhang7Xiaochun Wei8Pengcui Li9Shanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Department of Orthopedics, the Second Hospital of Shanxi Medical University, Taiyuan, ChinaShanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Department of Orthopedics, the Second Hospital of Shanxi Medical University, Taiyuan, ChinaShanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Department of Orthopedics, the Second Hospital of Shanxi Medical University, Taiyuan, ChinaShanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Department of Orthopedics, the Second Hospital of Shanxi Medical University, Taiyuan, ChinaShanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Department of Orthopedics, the Second Hospital of Shanxi Medical University, Taiyuan, ChinaShanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Department of Orthopedics, the Second Hospital of Shanxi Medical University, Taiyuan, ChinaShanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Department of Orthopedics, the Second Hospital of Shanxi Medical University, Taiyuan, ChinaShanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Department of Orthopedics, the Second Hospital of Shanxi Medical University, Taiyuan, ChinaShanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Department of Orthopedics, the Second Hospital of Shanxi Medical University, Taiyuan, ChinaShanxi Key Laboratory of Bone and Soft Tissue Injury Repair, Department of Orthopedics, the Second Hospital of Shanxi Medical University, Taiyuan, ChinaAims: To explore the novel molecular mechanisms of histone deacetylase 4 (HDAC4) in chondrocytes via RNA sequencing (RNA-seq) analysis. Methods: Empty adenovirus (EP) and a HDAC4 overexpression adenovirus were transfected into cultured human chondrocytes. The cell survival rate was examined by real-time cell analysis (RTCA) and EdU and flow cytometry assays. Cell biofunction was detected by Western blotting. The expression profiles of messenger RNAs (mRNAs) in the EP and HDAC4 transfection groups were assessed using whole-transcriptome sequencing (RNA-seq). Volcano plot, Gene Ontology, and pathway analyses were performed to identify differentially expressed genes (DEGs). For verification of the results, the A289E/S246/467/632 A sites of HDAC4 were mutated to enhance the function of HDAC4 by increasing HDAC4 expression in the nucleus. RNA-seq was performed to identify the molecular mechanism of HDAC4 in chondrocytes. Finally, the top ten DEGs associated with ribosomes were verified by quantitative polymerase chain reaction (QPCR) in chondrocytes, and the top gene was verified both in vitro and in vivo. Results: HDAC4 markedly improved the survival rate and biofunction of chondrocytes. RNA-seq analysis of the EP and HDAC4 groups showed that HDAC4 induced 2,668 significant gene expression changes in chondrocytes (1,483 genes upregulated and 1,185 genes downregulated, p < 0.05), and ribosomes exhibited especially large increases. The results were confirmed by RNA-seq of the EP versus mutated HDAC4 groups and the validations in vitro and in vivo. Conclusion: The enhanced ribosome pathway plays a key role in the mechanism by which HDAC4 improves the survival rate and biofunction of chondrocytes. Cite this article: Bone Joint Res 2023;12(7):433–446.https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.127.BJR-2022-0279.R2histone deacetylase 4rna sequencingribosomechondrocytegene mutationchondrocytesrnaswestern blottingmrnasgene expressionflow cytometryquantitative polymerase chain reactioncartilage tissuesrat modelsapoptosis |
| spellingShingle | Li Guo Hua Guo Yuanyu Zhang Zhi Chen Jian Sun Gaige Wu Yunfei Wang Yang Zhang Xiaochun Wei Pengcui Li Upregulated ribosome pathway plays a key role in HDAC4, improving the survival rate and biofunction of chondrocytes Bone & Joint Research histone deacetylase 4 rna sequencing ribosome chondrocyte gene mutation chondrocytes rnas western blotting mrnas gene expression flow cytometry quantitative polymerase chain reaction cartilage tissues rat models apoptosis |
| title | Upregulated ribosome pathway plays a key role in HDAC4, improving the survival rate and biofunction of chondrocytes |
| title_full | Upregulated ribosome pathway plays a key role in HDAC4, improving the survival rate and biofunction of chondrocytes |
| title_fullStr | Upregulated ribosome pathway plays a key role in HDAC4, improving the survival rate and biofunction of chondrocytes |
| title_full_unstemmed | Upregulated ribosome pathway plays a key role in HDAC4, improving the survival rate and biofunction of chondrocytes |
| title_short | Upregulated ribosome pathway plays a key role in HDAC4, improving the survival rate and biofunction of chondrocytes |
| title_sort | upregulated ribosome pathway plays a key role in hdac4 improving the survival rate and biofunction of chondrocytes |
| topic | histone deacetylase 4 rna sequencing ribosome chondrocyte gene mutation chondrocytes rnas western blotting mrnas gene expression flow cytometry quantitative polymerase chain reaction cartilage tissues rat models apoptosis |
| url | https://online.boneandjoint.org.uk/doi/epdf/10.1302/2046-3758.127.BJR-2022-0279.R2 |
| work_keys_str_mv | AT liguo upregulatedribosomepathwayplaysakeyroleinhdac4improvingthesurvivalrateandbiofunctionofchondrocytes AT huaguo upregulatedribosomepathwayplaysakeyroleinhdac4improvingthesurvivalrateandbiofunctionofchondrocytes AT yuanyuzhang upregulatedribosomepathwayplaysakeyroleinhdac4improvingthesurvivalrateandbiofunctionofchondrocytes AT zhichen upregulatedribosomepathwayplaysakeyroleinhdac4improvingthesurvivalrateandbiofunctionofchondrocytes AT jiansun upregulatedribosomepathwayplaysakeyroleinhdac4improvingthesurvivalrateandbiofunctionofchondrocytes AT gaigewu upregulatedribosomepathwayplaysakeyroleinhdac4improvingthesurvivalrateandbiofunctionofchondrocytes AT yunfeiwang upregulatedribosomepathwayplaysakeyroleinhdac4improvingthesurvivalrateandbiofunctionofchondrocytes AT yangzhang upregulatedribosomepathwayplaysakeyroleinhdac4improvingthesurvivalrateandbiofunctionofchondrocytes AT xiaochunwei upregulatedribosomepathwayplaysakeyroleinhdac4improvingthesurvivalrateandbiofunctionofchondrocytes AT pengcuili upregulatedribosomepathwayplaysakeyroleinhdac4improvingthesurvivalrateandbiofunctionofchondrocytes |